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Electrochemical sensor based on high-response glucose aptamer and preparation method thereof

A glucose and aptamer technology, applied in the field of electrochemical detection, can solve the problems of easy enzymatic degradation and poor stability, and achieve excellent sensitivity, large target response signal, and stable test signal

Pending Publication Date: 2021-10-26
武汉维尔博生物科技有限公司
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

Products at this stage often rely on glucose oxidase as the key element for detecting glucose. This type of sensor is prone to enzymatic degradation and other problems in a variable body fluid environment, and has poor stability.

Method used

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  • Electrochemical sensor based on high-response glucose aptamer and preparation method thereof
  • Electrochemical sensor based on high-response glucose aptamer and preparation method thereof
  • Electrochemical sensor based on high-response glucose aptamer and preparation method thereof

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preparation example Construction

[0045] The preparation method of the above-mentioned high-response glucose aptamer-based electrochemical sensor of the present invention comprises the following steps:

[0046] Step 1), making electrodes;

[0047] Step 2), configuring the aptamer solution according to claim 1;

[0048] Step 3), prepare the sensor, put the electrode of step 1) into the aptamer solution prepared in step 2) for reaction, then transfer it to the MCH solution for reaction, and take it out to prepare the detection sensor.

[0049] The nucleic acid aptamer of the present invention, in addition to being able to specifically bind to glucose, can also be combined with electrode materials through an infiltration reaction, which can greatly simplify the preparation process and difficulty of preparation, and has unexpected technical effects.

[0050] In a comparatively preferred embodiment of the present invention, in said step 1), the making of electrode comprises the following steps: select 2mm gold disk ...

Embodiment 1

[0057] Sensor based on the sequence of SEQ ID NO.1

[0058] The sensor of this embodiment is prepared by the following method, including the following steps:

[0059] Step 1), making gold disc electrodes.

[0060] Step 2), configure the aptamer solution based on the sequence of SEQ ID NO.1, the aptamer sequence is:

[0061] 5'-HO-C6-S-S-C6-ACGACCGTGTGTGTTGCTCTGTAACAGTGTCCATTGTCGT-MB-3';

[0062] The aptamer solution was reacted with TCEP solution to reduce the disulfide bond, and then diluted with 1×PBS buffer solution to obtain the desired aptamer reaction solution.

[0063] Step 3), prepare the sensor, put the gold disc electrode in step 1) into the aptamer reaction solution prepared in step 2) for reaction, then move it to the MCH solution for reaction, take it out, wash it, and dry it to prepare the present embodiment Examples of detection sensors.

Embodiment 2

[0065] Sensor based on the sequence of SEQ ID NO.2

[0066] The sensor of this embodiment is prepared by the following method, including the following steps:

[0067] Step 1), making a 2mm gold disc electrode.

[0068] Step 2), configure the aptamer solution based on the sequence of SEQ ID NO.2, the aptamer sequence is:

[0069] 5'-MB-TGCTGTTACCTGTGACAATGTCTCGTTGTGTGTGCCAG-HO-C6-S-S-C6-3';

[0070] The aptamer solution was reacted with TCEP solution to reduce the disulfide bond, and then diluted to 1 μM with 1×PBS buffer solution to obtain the desired aptamer reaction solution.

[0071] Step 3), prepare the sensor, put the gold disc electrode in step 1) into the aptamer reaction solution prepared in step 2), and react at room temperature; then move it to MCH solution, and react at 4°C; finally, take out , cleaning, and drying to prepare the detection sensor of this embodiment.

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Abstract

The invention discloses an electrochemical sensor based on a high-response glucose aptamer. A detection sensor comprises an electrode, and a glucose detection nucleic acid aptamer and a mercaptohexanol monomolecular layer are attached to the outside of the electrode; the glucose detection nucleic acid aptamer comprises an artificially synthesized nucleotide sequence capable of being specifically combined with glucose, a modification group and a methylene blue group, and the modification group is HO-C6-S-S-C6-; the artificially synthesized nucleotide sequence capable of being specifically combined with glucose is one of a sequence SEQ ID NO. 1, a sequence SEQ ID NO. 2, a sequence SEQ ID NO. 3 or a sequence SEQ ID NO. 4. The invention also provides a preparation method of the sensor. The nucleic acid aptamer is adopted as a recognition element, glucose can be detected in a whole blood environment with high sensitivity, participation of any enzyme is not needed, and the detection process is very convenient.

Description

technical field [0001] The invention relates to the technical field of electrochemical detection, in particular to an electrochemical sensor based on a high-response glucose aptamer and a preparation method thereof. Background technique [0002] Diabetes mellitus is a metabolic disease caused by insufficient insulin in the body caused by different pathogenesis and etiology, which leads to metabolic disorders of sugar, fat and protein, and severe hyperglycemia as the main clinical manifestation. Diabetes is divided into two main forms: type 1 diabetes (T1DM), which is an autoimmune disease that induces the destruction of insulin-producing pancreatic beta cells, resulting in an absolute lack of insulin; ② type 2 diabetes (T2DM), the main features It is insulin resistance, in which peripheral tissues cannot respond normally to insulin stimulation, and the body compensatoryly secretes excessive insulin to produce hyperinsulinemia to maintain blood sugar stability. [0003] Ther...

Claims

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Application Information

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Patent Type & Authority Applications(China)
IPC IPC(8): G01N27/327G01N27/49
CPCG01N27/3277G01N27/49
Inventor 李少光李辉朱满
Owner 武汉维尔博生物科技有限公司
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